Amahame ya Nucleic Acide Synthesis

Sintezike ya acide nucleic ikorwa hifashishijwe uburyo bukomeye bwa sialic amide triglyceride, aho 3 ′ iherezo rya ADN ryimuwe ku gice gikomeye kandi nucleotide ikongerwaho mu cyerekezo cya 3 ′ kugeza 5 ′ kugeza igihe igice cya ADN cyifuzwa gihurijwe hamwe. .Ibi bitandukanye na synthesis ya ADN ukoresheje ADN polymerase.

3 ′ iherezo ryibanze ryambere ryimuwe kuri CPG mugihe cya synthesis, 5′-OH yikibanza gikurikiraho irinzwe na di-p-tolyl trityl DMT, itsinda rya amino kuririndiro ririnzwe na acide benzoic na 3 ′ -OH noneho ikora hamwe na amino fosifate.1 shingiro ya 5 5′-OH ya 1 base na 3′-OH yikibanza gikurikiraho ikora fosifite triglyceride, hanyuma igahumeka hamwe na iyode kuri fosifate triglyceride, ikingira ikibanza cya kabiri 5′-OH ikurwaho na kongeramo aside ya dichloroacetic DM ikoresheje kongeramo urufatiro rukurikira, hanyuma nyuma ya synthesis ikingira ikingira kuri 5′-OH ikurwaho na acide idakomeye Igice cyaciwe na resin ikomeye hamwe na hydroxide ya amonium yibanze, umurinzi ukurwa mukibanza hamwe na hydroxide ya amonium yibanze munsi yubushyuhe, hydroxide ya amonium ikurwaho, igice kikaba cyumye, kandi aside nucleic isukurwa na chromatografiya yamazi cyangwa PAGE.

Intambwe za Synthesis ya Oligo

Gusiba

Uwiteka5 kurangiza DMT irinda itsinda ikurwaho wongeyeho igisubizo cya TCA.

Gukora

Kuvanga activate na oligonucleotide monomer kugirango ukore oligonucleotide monomer hagati.

Kubana

Itsinda rya hydroxyl ya 5-reaction ikora hamwe na oligonucleotide ikora hagati mugihe cyo kurwanya anti-generator reaction kugirango habeho isano ya fosifike triglyceride.

Gufata 

Kwiyongera kwa capping agent ikora acetylation capping reaction hamwe nitsinda ryinshi rya hydroxyl ridafite uruhare mubitekerezo bya kondegene.

Oxidation 

Kwiyongera k'umuti wa iyode urimo amazi bifata hamwe na fosifite idahindagurika hamwe na okiside kugirango bibeho aside fosifike ihamye triglyceride.

Cyanoethyl Cleavage

Igicuruzwa cya nucleic aside nyuma ya synthesis, hiyongereyeho igisubizo cya DEA kugirango ukureho itsinda rya cyanoethyl mumurongo wa fosifate triglyceride.

Byakuweho & Deprotection

Ibice bya acide nucleic biva mubice bitwara fonctionnement yongeramo hydroxide ya ammonium yibanze hanyuma bigashyuha kugirango bikureho amatsinda arinda imigozi ya hydrogène yibirindiro.